Indigenous enFnCas9 and TnpB-Based CRISPR Technology: Translational Applications in India

Abstract

AbstractCRISPR-based genome-editing technologies are progressing rapidly, outperforming conventionalCRISPR-Cas9 methods in medicinal and biotechnological applications. With unique structural andtranslational advantages, transposon-associated TnpB-based OMEGA systems and engineeredFrancisellanovicida Cas9 (enFnCas9) are two of the most promising next-generation genome-editingtechniques. EnFnCas9's enhanced selectivity, decreased off-target activity, and PAM-flexible editingmake it ideal for precision-medicine applications, including as ex vivo correction of monogenic disorders like sickle cell disease and β-thalassemia. TnpB-based editors, on the other hand, are ultra-compact RNA-guided nucleases made from IS200/IS605 transposons that provide easy dissemination, flexible TAM recognition, and rapid genome alteration in microbial and plant systems.Recent Indian innovations, such as the BIRSA-101 indigenous CRISPR therapy platform and the ICAR-CRRI-developed TnpB editing systems, demonstrate the emergence of a national genome-editing ecosystem focused on affordability, translational scalability, and reduced intellectual property dependence. This work explores the molecular architecture, mechanistic diversity, engineeringtechniques, delivery strategies, specificity profiling, translational applications, regulatory concerns,and pharmacological significance of the enFnCas9 and TnpB systems. Their uses in precisionmedicine, pharmacogenomics, medicinal plant engineering, gene-corrected cell therapy, andagricultural biotechnology are highlighted.